This research program focuses on Epstein-Barr virus Latent membrane protein 1 (LMP1) signaling, particularly the identification of molecular targets critical for LMP1-mediated NF-kB activation and target validation through genetic and chemical approaches. Specific objective 1 is to characterize LMP1 transmembrane interactions that are critical for LMP1 mediated NF-kB activation using reverse genetic, biochemical and fluorescence resonance energy transfer methodologies. Knowledge gained in these studies may enable interruption of transmembrane aggregation and signaling. Specific objective 2 focuses on identifying missing components of LMP1 TES1 and TES2 NF-kB activation by characterizing TRAF2, TRAF3, NIK, TRADD, TRAF6 and IRAKI associated proteins by tandem affinity pull-down experiments. Putative signaling components will be evaluated for their role in NF-kB activation by directed knock out using siRNA technology in B lymphocytes and epithelial cells. An siRNA based genetic screen will also be used to identify essential components of the NF-kB pathway. Specific objective 3 is a cell based screen for novel chemical inhibitors of LMP1 induced NF-KB that can serve as a chemical nucleus for novel therapeutics. The targets of such tool compounds will also be determined. NF- kB activity is required for EBV transformed B-lymphocyte survival. Thus inhibitors of LMP1 NF- kB activation will be useful for treating EBV associated diseases where LMP1 is expressed including lymphoproliferative disease in AIDS patients and transplant recipients, Hodgkin's disease, and Nasopharyngeal carcinoma. Given the importance of NF-kB activity in lymphocyte activation, growth, differentiation and survival, a tool compound might also provide direction for drug discovery applicable for allergy, transplantation, or auto immune disease.